Difference between revisions of "Part:BBa K3815005"

Revision as of 12:54, 20 October 2021

NOP1v-Mxe GryA intein-PT-linker-ELK16

Descriotion of this part

Targeted protein

This part is for the purification of NOP1v.This is made by adding a valine to the N-terminus of NOP1.The effect of it is the same as NOP1. We could not get NOP1 sufficiently when using BBa_K3815004. Then, we thought that considering N-end-rule, the N-terminus of NOP1 might have a negative effect on the recovery of peptide. Therefore, this part is made.

Sequence and Features

Purification system

In order to purify the peptide, we adopted ELK16 system that the past iGEM teams had not used. This part is composed of ELK16, Mxe Gyr intein, and PT linker. When this fused protein is produced, ELK16 self-assembles and precipitates. After that, taking this aggregate and adding DTT to it, the N terminal of Mxe GyrA intein is cut. Then, we can get the targeted protein.

We designed this part, however, we did not actually produce it with this part. We made this peptide with BBa_K3815010.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 120
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 120
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 120
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 120
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 120
Illegal NgoMIV site found at 553
1000
COMPATIBLE WITH RFC[1000]

@@ Line 8: / Line 8: @@
 Sequence and Features<br><br>
 <h3><font size="3">Purification system</font> </h3>
-In order to purify the peptide, we adopted ELK16 system that the past iGEM teams had not used. This part is composed of ELK16, Mxe Gyr intein, and PT linker. When this fused protein is produced, ELK16 self-assembles and precipitates. After that, taking this aggregate and adding DTT to it, the N terminal of Mxe GyrA intein is cut. Then, we can get the targeted protein.<br>
+In order to purify the peptide, we adopted ELK16 system that the past iGEM teams had not used. This part is composed of ELK16, Mxe Gyr intein, and PT linker. When this fused protein is produced, ELK16 self-assembles and precipitates. After that, taking this aggregate and adding DTT to it, the N terminal of Mxe GyrA intein is cut. Then, we can get the targeted protein.<br><br>
 We designed this part, however, we did not actually produce it with this part. We made this peptide with ''<partinfo>BBa_K3815010</partinfo>''.